Adult ; Female ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Regression Analysis ; Risk Factors ; Stress, Psychological ; psychology ; Surveys and Questionnaires ; Workplace ; psychology

OBJECTIVETo verify the interaction between secreted frizzled-related protein 2 (SFRP2) and osteoblast-specific factor 2 (OSF-2).

METHODSHA-tagged OSF-2 fusion protein recombinant vector pCMV-HA-OSF-2, which could express in mammal cells was constructed, then identified by enzyme-cutting and transfected into human kidney 293 (HK293) cells with or without Myc-SFRP2 recombinant eukaryotic expression vector pCMV-HA-SFRP2. The interaction between SFRP2 and OSF-2 was detected through coimmunoprecipitation and Western blotting.

RESULTSIn electrophoresis bath, target fragment of SFRP2 coding gene with 800 bp and target gene OSF-2 with 2500 bp could be seen respectively after enzyme-cutting, which showed that pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 were constructed successfully. No HA-OSF-2 expression was detected after pCMV-Myc-SFRP2 or pCMV-HA-OSF-2 transfection. Whereas, HA-OSF-2 expressed by Myc antibody immunoprecipitation after pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 co-transfection.

CONCLUSIONSHA-OSF-2 recombinant vector can express in mammal cells. Interaction exists between HA-OSF-2 and SFRP2.

Cell Line ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Genetic Vectors ; Humans ; Keloid ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Transfection

A high affinity polyclonal antibody-based enzyme linked immunosorbent assay (ELISA) was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1:5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 microg/ml, and the detection limit was 0.02 microg/ml for the assay. The overall recoveries and the coefficients of variation (CVs) were in the range of 82% to approximately 127% and 3.5% to approximately 8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol (ranging from 0.2 to 10 microg/ml) were detected by the ELISA and liquid chromatography (LC) method, and good correlations were obtained between the two methods (R(2)=0.9643). We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an alternative for the conventional LC method for zeranol in bovine urine.
Animals ; Calibration ; Cattle ; Chromatography, High Pressure Liquid ; Enzyme-Linked Immunosorbent Assay ; methods ; Indicator Dilution Techniques ; Zeranol ; immunology ; urine

OBJECTIVETo study the effects of specific small interfering RNA (siRNA) on Smoothened (Smo) gene expression and the proliferation and apoptosis of colorectal cancer LoVo cells.

METHODSThree different siRNAs (siRNA-1, siRNA-2, and siRNA-3, respectively) were transfected into LoVo cells via cationic liposome, and the changes of Smo mRNA level were determined using semi-quantitative RT-PCR 48 h after transfection. Flow cytometry and MTT assay were performed to assess the effect of the siRNAs on the proliferation and apoptosis of LoVo cells.

RESULTSForty-eight hours after Smo siRNA-1 transfection, Smo mRNA expression in LoVo cells decreased by about 63.56%, a reduction significantly greater than that in cells transfected with the other two siRNAs. The cell proliferation decreased significantly after Smo siRNA-1 transfection in comparison with the control cells, and 48 h after transfection, significantly higher apoptosis rate was observed in Smo siRNA-1-transfected cells than in the control cells.

CONCLUSIONSpecific siRNA can significantly decrease Smo mRNA expression and inhibit the proliferation while inducing apoptosis of LoVo cells.

Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; RNA, Small Interfering ; genetics ; Receptors, G-Protein-Coupled ; deficiency ; genetics ; Smoothened Receptor ; Time Factors ; Transfection

OBJECTIVETo investigate the expression of androgen receptor (AR) and hepatitis B virus X protein (HBx) in hepatocellular carcinoma (HCC), and analyze the relationship between AR and HBx expressions.

METHODSTumor tissues and peritumoral tissues of 83 HBV-associated HCC cases were investigated in this study. Fourteen cases of HBV-negative HCC and 13 cases of hemangioma peritumoral tissues were considered as control. AR and HBx mRNA levels were determined by quantitative fluorescence real-time RT-PCR and their protein levels were assayed by Western blot. The expression of AR and HBx proteins in tissues were examined with EnVision immunohistochemical staining. The methylation status of AR promoter was determined using methylation-specific PCR (MSP).

RESULTSBoth expression levels of AR mRNA and protein of the peritumoral tissues were significantly higher (0.17) than that of tumor tissues (0.09) in HBV-associated HCC (P < 0.01), but such a difference was not found in HBV-negative HCC (0.06 vs. 0.07, P > 0.05). The level of AR expression in peritumoral tissues was associated with tumor differentiation in HBV-associated HCC. AR mRNA and protein levels of peritumoral tissues in HBV-associated HCC were significantly higher than that in HBV-negative HCC and hemangioma (all P < 0.05). In the tumor tissues, HBV-associated HCC had significantly higher AR expression than HBV-negative HCC at mRNA level (P < 0.05), but not at protein level. Spearman rank correlation analysis showed that the AR mRNA or AR protein levels were positively correlated with HBx in both tumor and peritumoral tissues in HBV-associated HCC, but the expressions of AR and HBx were not associated with AR promoter methylation status. The relative expression levels of AR mRNA and protein in the HBV-associated peritumoral tissues were negatively correlated with tumor differentiation (r = -0.213, P < 0.05; r = -0.313, P < 0.05), the higher the AR expression, the poorer differentiation. But this correlation of AR mRNA and protein was not shown in the hepatocellular carcinoma tissues.

CONCLUSIONSHBx may enhance AR expression in HBV-associated HCC, but AR promoter demethylation maybe not been involved in its main mechanism. An increased AR expression is probably an early event during the development and progression of HBV-associated HCC, and AR expression in the peritumoral tissue is correlated with HBV-associated HCC differentiation. AR may play different roles in HBV-associated HCC and HBV-negative HCC.

Adult ; Aged ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; virology ; Cell Differentiation ; DNA Methylation ; Female ; Hemangioma ; metabolism ; Hepatitis B virus ; isolation & purification ; Humans ; Immunohistochemistry ; Liver ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Androgen ; genetics ; metabolism ; Trans-Activators ; metabolism

OBJECTIVETo investigate the expression and the role of secreted frizzled-related protein 2 (SFRP2) in the earlobe keloid and find a valid way to treat the keloid with gene therapy.

METHODSThe expression of SFRP2 mRNA and protein was tested with in situ hybridization and Western Blot Analysis method in the different period of earlobe keloid.

RESULTSThe SFRP2 mRNA and protein expression at the keloid edge was significantly high in 12 month group than in 3 or 6 month groups (P < 0.01), but not than in 24 month group. The SFRP2 expression started to decrease in the keloid center 12 month later (P < 0.01). The SFRP2 expression was always higher in edge than in center during all the period (P < 0.05, P < 0.01).

CONCLUSIONSThe results suggest that SFRP2 may play an important role in the development of keloid, especially at the keloid edge. The high SFRP2 expression in endothelial cells and surrounding tissue is also important. It may be a new way for gene therapy of keloid by decreasing the SFRP2 expression.

Adolescent ; Adult ; Ear ; Female ; Humans ; Keloid ; metabolism ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Young Adult

Aged ; Alcoholic Beverages ; Humans ; Hyponatremia ; etiology ; Male ; Substance Withdrawal Syndrome ; complications

OBJECTIVETo study difference in occupational stress between men and women commercial workers in a supermarket in Yinchuan, Ningxia.

METHODSTotally, 679 commercial workers in a supermarket were investigated with questionnaire of occupational stress indicator (OSI), matched on age, length of service, educational level, marital status and type of work.

RESULTSScore of occupational stress factors, relationships, home/work balance and organizational atmosphere in women commercial workers was 143.48, 30.86, 20.82 and 15.16, respectively, obviously higher than that in men, with 134.89, 28.61, 18.75 and 13.93, respectively. Score of psychological health and satisfaction in women was 39.86 and 14.82, respectively, lower than that in men, with 43.84 and 17.66, respectively, which indicate that occupational stress in women was more severe with a more stressful psychological reaction than in men. Stepwise regression analysis showed that the main predicting factor for job satisfaction was personal relationships in women, and organizational atmosphere, managerial role and workload in men. Those for psychological health was control strategy and organizational atmosphere in women, and organizational atmosphere and recognition in men, those for physical illness was workload in women and support strategy and physical exercises in men, and those for stress level was support strategy in women and coping strategy in men.

CONCLUSIONSWomen commercial workers experienced much more stress, with more severe stress reaction in their work, than men did. The main factors affecting occupational stress reaction and level of stress in women and men were not quite similar.

Adaptation, Psychological ; Adult ; China ; epidemiology ; Commerce ; Female ; Humans ; Interpersonal Relations ; Job Satisfaction ; Male ; Sex Factors ; Stress, Psychological ; epidemiology ; psychology ; Surveys and Questionnaires ; Workload

The latest research progress on quantitative determination methods of main active components-lignans from Schisandra chinensis and its preparations has been summarized, such as spectrophotometry, thin-layer chromatography scanning, high performance liquid chromatograpy, gas chromatography-mass spectrometry and capillary electrochromatography. The characteristics and application areas of every analytical method have also been stated. It offers reference on quality control of crude drug and its preparations of S. chinensis.
Capsules ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Thin Layer ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fruit ; chemistry ; Gas Chromatography-Mass Spectrometry ; Lignans ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Schisandra ; chemistry